4916 results about "Target gene" patented technology

The invention relates to a high-activity recombinant XVII type collagen as well as a rapid preparation method and application thereof, and belongs to the technical field of bioengineering. The amino acid sequence of the high-activity recombinant XVII type collagen is as shown in SEQ ID NO. 1, and the nucleotide sequence of the high-activity recombinant XVII type collagen is as shown in SEQ ID NO. 2. The invention also provides a preparation method and application of the recombinant XVII type collagen. The high-activity recombinant XVII type collagen has good biological activity, has good effects of promoting cell adhesion, cell proliferation and cell migration, and can be compounded with other proteins to form raw materials of tissue engineering products, cosmetics, health care products or medicines. The high-activity recombinant XVII type collagen gene is integrated to different positions of a yeast genome through three different vectors, and the copy number of a target gene is increased, so that the expression quantity of the recombinant XVII type collagen is remarkably increased, and the yield of the high-activity recombinant XVII type collagen is effectively increased.

The present invention relates to transmissible elements (e.g. conjugative plasmids, phage and phagemid) which are engineered to express molecules of interest (MOIs) and a pathway modification system (PMS) which is able to introduce mutations in target endogenous genes in a recipient bacterium, or is able to produce nucleic acids which inhibit or increase expression of target endogenous genes in a recipient bacterium, or is able to express peptide inhibitor molecules which inhibit a target endogenous protein in a recipient bacterium. The invention relates to host cells containing the transmissible elements, to pharmaceutical compositions containing such transmissible elements or host cells, and their use in the treatment of various diseases.

A modification template for improving the inhibitory effect of a double-stranded oligonucleotide on a target gene, a combination thereof and the use thereof. The double-stranded oligonucleotide is an siRNA and contains a sense strand and an antisense strand, and each nucleotide of the sense strand and antisense strand is a modified nucleotide. The double-stranded oligonucleotide is modified with a special modification template, and has a significantly improved inhibition rate on a target gene compared with a basic sequence.

The invention belongs to the technical field of RNA biological prevention and treatment, and particularly relates to dsRNA for preventing and treating crop anthracnose and application of the dsRNA. The dsRNA is targeted inositol phosphatidylceramide (IPC) synthetase coding gene dsRNA, and is obtained by transcription of a part of sequence (SEQ ID No.1) of an inositol phosphatidylceramide (IPC) synthetase coding gene CaAUR1, the gene segment is a segment of a target gene for preventing and treating crop anthracnose, and the dsRNA obtained by transcription can obviously reduce pathogenicity of mango anthracnose bacteria, and has the effects of keeping mango fresh, preventing and treating anthracnose. And the compound can be used for preventing and treating the crop anthracnose or preparing products for preventing and treating the crop anthracnose.

Provided are a double-stranded oligonucleotide agent and uses. The double-stranded oligonucleotide agent comprises a specific cleavage site for an endonuclease, wherein the cleaved double-stranded oligonucleotide agent is capable of inhibiting expression of the target gene via RNA interference (RNAi).

The invention discloses a site for stably expressing protein in a CHO cell gene NW023276806.1 and application of the site, and belongs to the technical field of biological genes. The site belongs to a fixed position in a CHO cell genome, different protein genes are introduced based on a micro-homologous end connection mechanism through a CRISPR / Cas9 tool, and stable expression is carried out. By adopting a site-specific integration method, a target gene is integrated to a stable expression area in a site-specific manner, repeated high-expression monoclonal screening is effectively avoided, and an MMEJ mechanism is introduced to integrate a donor fragment, so that the research and development time for constructing a stable expression cell strain in biological pharmacy can be effectively shortened, and the cost is reduced.

The invention discloses a causal gene regulation network inference method based on a directed graph attention network, and belongs to the technical field of graph representation learning and system biology. According to the method, an initial directed causal graph is constructed by using prior regulation and control information, expression data is used as initial features of nodes in the graph, and then two directed graph attention networks are used for respectively learning embedded expressions of genes as source nodes and target nodes in the directed causal graph; meanwhile, learning feature information of genes implied in the expression data by using a residual network; and finally, calculating the probability of the potential directed regulation relationship based on transcription factor embedding, target gene embedding and self-embedding of the gene. According to the method, the gene regulation network is learned and modeled from the perspective of the directed regulation relationship, and the inference accuracy of the potential directed regulation relationship is improved.

The invention discloses an intercellular signal transduction inference method and device based on optimal transmission of a knowledge graph, equipment and a medium, and relates to the technical field of genetic engineering. The source gene expression data of different cells in the source cell type and the target gene table data of different cells in the target cell type are found according to the cell type database. On the basis of the ligand-receptor interaction knowledge graph, obtaining various pathways with an interaction relationship between the source cell type and the target cell type, and constructing a pathway set; and with the aim of minimizing the total transmission cost, determining an optimal transmission cost matrix of each path in the path set by adopting an scKGOT algorithm. According to the method, the intercellular channel can be more conveniently deduced, and the intercellular communication can be deduced by accurately simulating the intercellular ligand-receptor signal channel.

The invention relates to the technical field of bioinformatics, in particular to a method for identifying an elncRNAs regulation target gene in a single cell based on single cell ATAC-seq. According to the method provided by the invention, a regulatory network of an elncRNAs-target gene on a single cell whole genome is identified by utilizing chromatin co-accessibility based on scATAC-seq data. Meanwhile, the method is applied to immune cell differentiation, and the elncRNAs-target gene regulation dynamic state of immune cell differentiation is accurately recognized. In addition, the biological functions of the elncRNAs in different cell types are further analyzed. The method provided by the invention can help related researchers in the field of biomedicine to more elaborately and accurately predict the regulation effect and function of the elncRNAs under the single cell resolution.

The invention belongs to the technical field of immunology and in vitro diagnosis, and particularly relates to a rabbit-derived recombinant monoclonal antibody for specific recognition of type II grass carp reovirus VP4 protein, an eukaryotic expression method and application. According to the invention, a target gene S6 is transfected to an HEK293 cell for expression and purification, a New Zealand white rabbit is immunized in cooperation with a Freund's adjuvant, and the rabbit-derived recombinant monoclonal antibody capable of specifically recognizing the GCRV-II VP4 protein is obtained by using ELISA, single B cell screening and eukaryotic expression technologies. The antibody has strong specificity, provides support for further establishment of a specific II-type grass carp reovirus diagnosis technology, and has great application value in development of GCRV-II related scientific research.

The invention provides a siRNA gene inhibition efficiency screening model construction and training method, a siRNA gene inhibition efficiency screening device and a siRNA gene inhibition efficiency screening method. The construction and training method comprises the following steps: acquiring first sequence information of target mRNA and second sequence information of siRNA; obtaining first space image information of the target mRNA and second space image information of the siRNA; constructing a siRNA gene inhibition efficiency screening model, and training a first neural network model by using the first sequence information and the second sequence information to extract sequence features; training a second neural network model by using the first spatial image information and the second spatial image information to extract spatial structure features; and fusing the sequence features and the spatial structure features, and training a third neural network model by using the fused features. Therefore, the accuracy of screening the siRNA with the inhibition efficiency on the target gene can be improved.

The invention provides recombinant IV-type collagen as well as a preparation method and application thereof, and relates to the technical field of biochemical engineering. The preparation method of the recombinant IV-type collagen comprises the following steps: obtaining a codon from amino acid of the recombinant IV-type collagen, further obtaining a target gene, and synthesizing the target gene onto an expression vector to obtain a plasmid; transferring the plasmid into an expression host, and screening a positive transformant to obtain an expression strain; and culturing the expression strain, and expressing the target protein to obtain the crude recombinant IV-type collagen. The high-purity recombinant IV-type collagen is prepared through a genetic engineering technology, and the recombinant IV-type collagen can significantly down-regulate inflammatory factors and reduce inflammation; heat injury can be reduced by up-regulating heat shock protein; after heat injury, the recombinant IV-type collagen can obviously lower metal matrix protease of heat injury cells, and also can obviously promote gene expression of type I collagen and type III collagen, improve skin elasticity and moisture content and delay skin aging.

The invention belongs to the field of biological medicines, and particularly relates to a modified double-stranded oligonucleotide, and a conjugate and a composition thereof. The double-stranded oligonucleotide provided by the invention effectively enhances the activity of the double-stranded oligonucleotide for inhibiting the expression of the target gene by performing specific modification on the nucleotide at a specific site, can efficiently inhibit the expression of the target gene in both in-vitro and in-vivo experiments, and surprisingly, the double-stranded oligonucleotide provided by the invention has a good application prospect. The modification on the double-stranded oligonucleotide can be generally used for siRNA designed for various different target genes, and the inhibition rate of the siRNA on the target genes can be effectively improved.

The invention provides a plant salt tolerance related transcription factor TaVOZ1 as well as a coding gene and application thereof, and belongs to the technical field of functional genes. The TaVOZ1 is related to plant salt tolerance and has an amino acid sequence as shown in SEQ ID No. 1, and a gene for coding the protein has a nucleotide sequence as shown in SEQ ID No. 2. The plant salt tolerance can be regulated by regulating the content or activity of the protein or the expression quantity of the gene, for example, when the gene is over-expressed, the plant salt tolerance is enhanced, and when the gene is knocked down, the salt tolerance is weakened. The TaVOZ1 disclosed by the invention can be combined with 5 '-CTTCT-3'or a reverse complementary sequence 5'-AAGAAG-3 'in a downstream target gene promoter to activate the expression of a stress response gene so as to improve the salt tolerance of a plant, so that the TaVOZ1 can be used for culturing a salt-tolerant wheat variety.

The present invention relates to T lymphocytes targeting CLD18A2 and a preparation method and application thereof. The inventors have successfully found a target gene CLD18A2 suitable for developing CAR T cells from a number of tumor-related genes for the first time, and successfully prepared CAR T cells targeting CLD18A2, thereby providing a new treatment method for pancreatic cancer, gastric cancer and other tumors.

The invention discloses a modification template for improving the target gene inhibition effect of double-stranded oligonucleotide and a combination and application of the modification template, the double-stranded oligonucleotide is siRNA and contains a positive-sense strand and an antisense strand, and each nucleotide of the positive-sense strand and the antisense strand is a modified nucleotide. The double-stranded oligonucleotide is modified by a special modification template, and compared with a basic sequence, the inhibition rate of the double-stranded oligonucleotide to a target gene is remarkably improved.

The present invention discloses a recombinant sheep β-defensin 2 protein expressed by Pichia pastoris and its application. The recombinant sheep β-defensin 2 gene designed according to the CDS of the sheep β-defensin 2 gene is integrated into the genome of Pichia pastoris GS115, thereby realizing the secretory expression of the target gene on Pichia pastoris. The recombinant sheep β-defensin 2 protein expressed by Pichia pastoris eukaryotic cells has a strong antibacterial effect and has broad application prospects in the development of new antibiotic alternative products and antimicrobial peptides for sheep feed.

The invention discloses a method for constructing a prediction model of pan-apoptosis related biomarkers of asthma based on a convolutional neural network algorithm and application, and the method comprises the steps: firstly obtaining related data from a database, and determining characteristic pan-apoptosis related genes PDGs in asthma through an SVM support vector machine, LASSO regression analysis and RF random forest regression; the method comprises the following steps: establishing an lncRNA-miRNA-mRNA network by using interaction STRING and Cytoscape software, constructing a prediction model of the pan-apoptosis related biomarkers of asthma through a convolutional neural network, and analyzing the diagnosis accuracy of potential target genes through an ROC curve. 10 characteristic PDGs playing an important role in asthma are finally obtained, the difference of the genes between a normal group and an asthma group is proved, and the enrichment pathway and immune cell infiltration of the genes have a certain relation with the occurrence and development of asthma, so that a prediction model of the pan-apoptosis related biomarkers of asthma is constructed, and the prediction model is used for predicting the pan-apoptosis related biomarkers of asthma. And a new research direction is provided for studying the pathogenesis and treatment of asthma.

The invention provides an intracellular mycobacterium tuberculosis in-situ nucleic acid detection method based on CRISPR / Cas and application, and the method comprises the following steps: S1, designing crRNA and a primer pair of a mycobacterium tuberculosis target gene sequence, and synthesizing a single-stranded DNA fluorescent probe; s2, preparing a standard solution containing a target gene sequence, and incubating with a CRISPR / Cas12a system; s3, extracting nucleic acids of different types of bacteria, uniformly mixing the nucleic acids of different types of bacteria with the CRISPR / Cas12a system, and incubating; s4, incubating an attenuated strain of mycobacterium tuberculosis and the DNA fluorescent probe, infecting macrophages, and incubating to obtain infected cells; s5, electrically transferring a CRISPR / Cas12a system into the infected cells in S4 by utilizing an electroporation technology, incubating, and detecting the intensity of a fluorescence signal; and S6, performing quantitative and positioning analysis on the detected fluorescence signal to complete detection, and directly introducing a CRISPR / Cas12a system into the macrophage by adopting an electroporation technology to realize intracellular in-situ detection, thereby avoiding the operation complexity of a nucleic acid extraction link and the loss of detection sensitivity in the prior art.

This disclosure describes methods, non-transitory-computer readable media, and systems that can simultaneously determine estimated methylation-level values for cytosine bases and genotype calls for a target genomic sample. The disclosed system can utilize a Bayesian method on a target genomic sample's nucleotide-read data to generate estimated methylation-level values that indicate genomic coordinates at which the target genomic sample comprises a reference cytosine base or a nucleobase that could be called as a cytosine. The disclosed system can estimate methylation-level values based on prior genotype probabilities and observed nucleobases at a genomic coordinate from a read pileup of a target genomic sample. Based on the estimated methylation-level values and base-call-quality metrics, the disclosed system may generate posterior genotype probabilities for the genomic sample at the genomic coordinate. Based on the posterior genotype probabilities, the disclosed system can generate a genotype call for the target genomic sample.

The invention relates to the field of biological medicine, in particular to a nucleoside analogue and application thereof to reduction of off-target activity of oligonucleotide drugs. The nucleoside analogue provided by the invention can be used for preparing oligonucleotide drugs, and can be coupled to a seed region of siRNA, so that the specificity of siRNA is efficiently improved, and the silence effect of a target gene can be maintained while the off-target effect of siRNA is reduced.

The invention relates to the technical field of high-throughput sequencing, and discloses a kit and a library construction method and a target variation detection and analysis method thereof, the kit comprises a pre-library construction reagent and a target region liquid phase hybridization capture reagent; wherein the pre-library construction reagent comprises a repair reagent, a joint, a connection reagent, an amplification reagent and purified magnetic beads; the target area liquid phase hybridization capture reagent comprises a hybridization reagent, a cleaning reagent, an amplification reagent, a purification magnetic bead, a target gene probe and a CNV / LOH probe. The invention provides a rapid, accurate and low-cost construction method of a library containing a targeting gene probe and a CNV / LOH probe and application of the library, through one-time library construction and sequencing, the advantages of a high-throughput targeting sequencing technology and a CNV-Seq technology are integrated; multi-gene point mutation, insertion and deletion, gene copy number variation, large-fragment copy number variation and heterozygosity deletion can be detected at the same time.

The invention provides a primer probe group for simultaneously detecting American strains and European strains of porcine reproductive and respiratory syndrome viruses, a detection method and application, and particularly belongs to the technical field of porcine pathogen detection products. The invention provides a primer probe group for simultaneously detecting American strains and European strains of porcine reproductive and respiratory syndrome viruses. The primer probe group comprises universal forward primers PRRSV1-F and PRRSV2-F for detection, reverse primers PRRSV1-R and PRRSV2-R for detection and a universal probe PRRSV-P. A target gene detected by the primer probe group is a 3'non-coding region gene (3 'UTR) of a porcine reproductive and respiratory syndrome virus genome, and the primer probe group can realize efficient detection of American strains and European strains of the porcine reproductive and respiratory syndrome virus, and is simple and convenient in detection, high in sensitivity, good in specificity, high in speed, low in false positive and capable of realizing high-throughput detection.

The invention discloses a coconut strong promoter, a plant expression vector and application. The nucleotide sequence of the promoter is as shown in SEQ ID NO. 1. Compared with a known promoter, the promoter provided by the invention can drive high-efficiency expression of related genes, and is a strong promoter. The invention also relates to a plant expression vector containing the strong promoter and application of the plant expression vector in promoting expression of a target gene.

The invention provides a primer probe composition and a kit for detecting a Databean virus and application, and relates to the technical field of biological detection, the primer probe composition comprises a primer probe group aiming at a Databean virus S gene and a primer probe group aiming at a Databean virus L gene, the Databean virus L and S genes are used as amplification target genes for design, and the kit is used for detecting the Databean virus. The primer probe group aiming at the S gene of the Databu virus and the primer probe group aiming at the L gene of the Databu virus are used as the primer probe composition, so that multi-gene target amplification is realized, and the positive detection rate reaches 100% and is higher than the positive detection rate of single-gene amplification; the lowest detection limit is up to 200 copies / mL; the quantification limit reaches 500 copies / mL, and the quantification linear range is 5.0 * 10 < 2 >-1.0 * 10 < 8 > copies / mL. The technical problem that in the prior art, a high-sensitivity and high-accuracy multiplex qPCR detection method for the nucleic acid of the copyda virus is lacked is solved.

The invention provides a beauveria bassiana engineering strain Bb-dsMucin for expressing dsRNA (double-stranded ribonucleic acid) as well as a construction method and application of the beauveria bassiana engineering strain Bb-dsMucin. The beauveria bassiana is a beauveria bassiana QSE-F1 strain, and the strain can be colonized at the tomato leaves. The dsRNA can target a botrytis cinerea key gene BcMucin, and the expression of the BcMucin gene is silenced in an RNAi (Ribonucleic Acid Interference) mode. According to the engineering strain, a carrier containing a target gene sequence is transferred into a beauveria bassiana QSE-F1 strain by utilizing an ATMT technology, and the engineering strain Bb-dsMucin capable of synthesizing dsRNA of targeted BcMucin is obtained. The invention also provides an application of the engineering strain Bb-dsMucin, and the application is that the engineering strain Bb-dsMucin can interfere with the expression of a target gene BcMucin, so that the botrytis cinerea can be effectively prevented and treated. Experiments prove that the provided engineering strain Bb-dsMucin can be used for preparing biological agents for preventing and treating gray mold and has a good market application prospect.

The invention belongs to the technical field of bioengineering, and provides a recombinant XVII type collagen as well as a preparation method and application thereof. A target gene sequence is obtained through bioinformatics predictive analysis and integrated design on the basis of a human XVII type collagen sequence, codon optimization is carried out, a T7 promoter in a plasmid vector is optimized, an engineering strain of the high-hydrophobicity humanized XVII type collagen is prepared, and the high-hydrophobicity humanized XVII type collagen is obtained. The strain is subjected to culture and inducible expression to obtain high-yield recombinant XVII type collagen. The recombinant XVII type collagen disclosed by the invention is reasonable in sequence design, large in expression quantity, simple in preparation method and stable in heredity, and rapid separation of the collagen can be realized in a production and preparation process by utilizing high hydrophobicity.

The invention provides a cell specific transcription factor regulatory network analysis method and a visualization platform, and relates to the technical field of bioinformatics, the method comprises the following steps: constructing a gene regulatory network through a GRNBoost2-cisTarget-AUCell-Cell GRN workflow based on a transcription factor in combination with a motif database; screening a direct regulation relationship in combination with the database, and calculating an activity score of a regulator in each cell; based on activity scores and cell type annotation results, grouping the single cell data by using a unified manifold approximation and projection (UMAP) dimensionality reduction method and a Leiden clustering algorithm, and displaying the following results through an interactive visualization tool: a cell clustering UMAP graph, performing color marking according to cell types; a UMAP graph and a heat map of transcription factor regulator activity; according to the visual map of the gene regulation and control network, transcription factors and target genes are distinguished through node shapes, and regulation and control relations are marked through line weights and colors. According to the invention, an accurate regulation and control network can be provided.

The invention discloses an application of a soybean SDR family gene GmGAE47 in cultivation. The invention discloses a cultivated soybean GmGAE47 protein coding gene GmGAE47. The nucleotide sequence of the cultivated soybean GmGAE47 protein coding gene is SEQ ID NO. 1. The constructed plant overexpression vector Pba002-GmGAE47 is subjected to heterologous expression in a wild type of arabidopsis thaliana, and it is found that a transgenic plant changes in the aspects of plant height, thousand seed weight and grain width characters. The gene can be introduced into a plant as a target gene, and the plant height and yield of the transgenic plant are regulated and controlled through the expression of the GmGAE47 gene. Therefore, the cultivated soybean GmGAE47 protein coding gene GmGAE47 disclosed by the invention can be applied to the aspects of regulating the plant height, increasing the thousand seed weight, the grain width and the like through genetic engineering.

The present invention provides a model or tool for predicting the editing efficiency of sgRNA in a CRISPR / Cas gene editing system, in particular a CRISPR / dCas epigenetic editing system, a training and prediction method thereof, and a related computer system, computer storage medium, and application. In particular, one or more sgRNA and target gene related epigenetic features are added to an input of the model.